Guo Chen, Huang Tao, Wang Qing-Hai, Li Hong, Khanal Aashish, Kang En-Hao, Zhang Wei, Niu Hai-Tao, Dong Zhen, Cao Yan-Wei
1Department of Urology, The Affiliated Hospital of Qingdao University, No. 59 Haier Road, Qingdao, 266071 Shandong China.
2Department of Pathology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong China.
Cancer Cell Int. 2019 Jun 28;19:170. doi: 10.1186/s12935-019-0889-8. eCollection 2019.
The Warburg effect demonstrates the importance of glycolysis in the development of primary and metastatic cancers. We aimed to explore the role of monocarboxylate transporter 1 (MCT1) and MCT4, two essential transporters of lactate, in renal cancer progression during cancer-endothelial cell co-culturing.
Renal cancer cells (786-O) and human vascular endothelial cells (HUVECs) were single-cultured or co-cultured in transwell membranes in the presence or absence of a MCT-1/MCT-4 specific blocker, 7ACC1. Cell proliferation was evaluated with the CCK-8 kit, while cell migration, after a scratch and invasion in transwell chambers, was evaluated under a microscope. Real-time qPCR and western blot were employed to determine the mRNA and protein levels of MCT1 and MCT4, respectively. The concentration of lactic acid in the culture medium was quantified with an l-Lactic Acid Assay Kit.
786-O cells and HUVECs in the co-culturing mode exhibited significantly enhanced proliferation and migration ability, compared with the cells in the single-culturing mode. The expression of MCT1 and MCT4 was increased in both 786-O cells and HUVECs in the co-culturing mode. Co-culturing promoted the invasive ability of 786-O cells, and markedly increased extracellular lactate. Treatments with 7ACC1 attenuated cell proliferation, migration, and invasion, and down-regulated the levels of MCT1/MCT4 expression and extracellular lactate.
The Warburg effect accompanied with high MCT1/MCT4 expression in the cancer-endothelial microenvironments contributed significantly to renal cancer progression, which sheds new light on targeting MCT1/MCT4 and glycolytic metabolism in order to effectively treat patients with renal cancers.
瓦伯格效应表明糖酵解在原发性和转移性癌症发展中的重要性。我们旨在探讨乳酸的两种重要转运体——单羧酸转运蛋白1（MCT1）和MCT4在肾癌细胞与内皮细胞共培养过程中对肾癌进展的作用。
肾癌细胞（786 - O）与人血管内皮细胞（HUVECs）在存在或不存在MCT - 1/MCT - 4特异性阻滞剂7ACC1的情况下，在Transwell膜中进行单培养或共培养。用CCK - 8试剂盒评估细胞增殖，而在划痕和Transwell小室侵袭后，在显微镜下评估细胞迁移。采用实时定量PCR和蛋白质印迹法分别测定MCT1和MCT4的mRNA和蛋白质水平。用l - 乳酸检测试剂盒定量培养基中乳酸的浓度。
与单培养模式下的细胞相比，共培养模式下的786 - O细胞和HUVECs表现出显著增强的增殖和迁移能力。共培养模式下786 - O细胞和HUVECs中MCT1和MCT4的表达均增加。共培养促进了786 - O细胞的侵袭能力，并显著增加了细胞外乳酸。用7ACC1处理可减弱细胞增殖、迁移和侵袭，并下调MCT1/MCT4表达水平和细胞外乳酸。
癌症 - 内皮微环境中伴随高MCT1/MCT4表达的瓦伯格效应显著促进了肾癌进展，这为靶向MCT1/MCT4和糖酵解代谢以有效治疗肾癌患者提供了新的思路。
